CAJ | 학술논문

目的研究沉默信息调节因子1 (SIRT1)基因沉默对间充质干细胞(MSCs)受照后Nod样受体蛋白3(NLRP3)和IL-1β表达的影响,探讨SIRT1激活剂——白藜芦醇的辐射防护作用及其机理.方法将MSCs分为空白对照组、单纯照射组、RNA干扰组、白藜芦醇组和RNA干扰+白藜芦醇组.采用酶联免疫吸附测定、Western blot和RT-PCR等方法检测IL-1β、SIRT1和NLRP3在蛋白和mRNA水平的表达.结果辐射可导致MSCs细胞外IL-1β分泌水平明显升高,给予白藜芦醇后,细胞IL-1β分泌水平较单纯照射组显著下降(t=21.68,P＜0.01),NLRP3和IL-1β mRNA水平较单纯照射组明显降低(t=14.44,P＜0.01;t=12.35,P＜0.01),SIRT1基因沉默后,NLRP3和IL-1β的mRNA水平回升至单纯照射组水平(t=14.86,P＜0.01;t=11.12,P＜0.01),即使再给予白藜芦醇,NLRP3和IL-1β的mRNA水平仍明显高于白藜芦醇组(t=11.31,P＜0.01;t=10.54,P＜0.01).结论 SIRT1基因沉默减弱了白藜芦醇对辐射诱导的NLRP3和IL-1β的抑制作用,说明白藜芦醇可能通过激活SIRT1、抑制NLRP3、降低IL-1β的表达,从而减轻辐射引起的细胞损伤.
목적 연구침묵신식조절인자1 (SIRT1)기인침묵대간충질간세포(MSCs)수조후Nod양수체단백3(NLRP3)화IL-1β표체적영향,탐토SIRT1격활제——백려호순적복사방호작용급기궤리.방법 장MSCs분위공백대조조、단순조사조、RNA간우조、백려호순조화RNA간우+백려호순조.채용매련면역흡부측정、Western blot화RT-PCR등방법검측IL-1β、SIRT1화NLRP3재단백화mRNA수평적표체.결과 복사가도치MSCs세포외IL-1β분비수평명현승고,급여백려호순후,세포IL-1β분비수평교단순조사조현저하강(t=21.68,P＜0.01),NLRP3화IL-1β mRNA수평교단순조사조명현강저(t=14.44,P＜0.01;t=12.35,P＜0.01),SIRT1기인침묵후,NLRP3화IL-1β적mRNA수평회승지단순조사조수평(t=14.86,P＜0.01;t=11.12,P＜0.01),즉사재급여백려호순,NLRP3화IL-1β적mRNA수평잉명현고우백려호순조(t=11.31,P＜0.01;t=10.54,P＜0.01).결론 SIRT1기인침묵감약료백려호순대복사유도적NLRP3화IL-1β적억제작용,설명백려호순가능통과격활SIRT1、억제NLRP3、강저IL-1β적표체,종이감경복사인기적세포손상.
Objective To investigate the effect of silence information regulator-1 (SIRT1) on NLRP3 and IL-1β production in mesenchymal stem cells (MSCs) exposed to radiation and to explore the SIRT1 role against radiation.Methods MSCs were divided into control, irradiation, RNAi, resveratrol, and RNAi+resveratrol group.IL-1β, SIRT1, and Nod-like receptor protein 3 (NLRP3) expressions were detected using enzyme-linked immunosorbent assay (ELISA), Western blot analysis, and real-time PCR assay.Results Extracellular IL-1β secretion induced by radiation was increased significantly.After resveratrol treatment, the levels of IL-1β secretion decreased compared with those of the radiation group (t=21.68, P＜0.01).The mRNA level of NLRP3 and IL-1β also decreased compared with that of the radiation group (t=14.44, P＜0.01;t=12.35, P＜0.01).SIRT1 knockdown significantly suppressed resveratrol-mediated anti-inflammatory activity(t=14.86, P＜0.01;t=11.12, P＜0.01).The mRNA level of NLRP3 and IL-1β remained higher in the RNAi+resveratrol group than those in the resveratrol group (t=11.31, P＜0.01;t =10.54, P＜0.01).Conclusions SIRT1 gene silencing can suppress NLRP3 and IL-1β expression induced by radiation.Thus, resveratrol may alleviate cellular damage induced by radiation through activation of SIRT1, inhibition of NLRP3, and decrease in IL-1β expression.